magistrsko delo
Povzetek
Keratinski filamenti so glavni strukturni element v epitelijskih celicah. V celicah vzdržujejo obliko in trdnost ter zagotavljajo fizično odpornost na mehanske obremenitve. Opisanih je bilo več različnih mutacij v keratinih, ki vodijo v različne tipe bolezni. Za mutacije v keratinih 5 in 14 je znano, da povzročijo nastanek bulozne epidermolize simpleks (EBS), bolezni krhkosti kože. V magistrskem delu smo preučevali točkovno mutacijo K14 R125P, ki povzroča hudo obliko EBS. Tipičen fenotipski pojav ob mutaciji je nastanek agregatov, ki pa še ni natančno razložen. Z uporabo optične pincete smo določili togost keratinocitov z vstavljenim konstruktom dodatne kopije K14, z mutacijo R125P, za različne količine mutiranega keratina. Ugotovili smo, da se razlika v togosti pojavi zgolj pri celicah z najnižjim deležem mutiranega keratina, pri katerih smo opazili tudi največ celic z agregati. Agregati so dinamičen organel, ki ga celica lahko reciklira s pomočjo proteosomalnega sistema, kot smo dokazali z njegovo inhibicijo in prenosom po Westernu. Z matematičnim modeliranjem smo razložili tudi nastanek agregatov – z asimetrično vezavo, pri kateri sodelujeta tako keratin divjega tipa kot mutirani keratin, kasneje pa agregati rastejo ob dodajanju predvsem mutiranega keratina.
Ključne besede
koža;epidermis;keratinociti;keratin 14;intermediarni filamenti;agregati;bulozna epidermoliza simpleks;magistrska dela;
Podatki
| Jezik: |
Slovenski jezik |
| Leto izida: |
2021 |
| Tipologija: |
2.09 - Magistrsko delo |
| Organizacija: |
UL FKKT - Fakulteta za kemijo in kemijsko tehnologijo |
| Založnik: |
[T. Sorčan] |
| UDK: |
576(043.2) |
| COBISS: |
82715907
|
| Št. ogledov: |
223 |
| Št. prenosov: |
23 |
| Ocena: |
0 (0 glasov) |
| Metapodatki: |
|
Ostali podatki
| Sekundarni jezik: |
Angleški jezik |
| Sekundarni naslov: |
Determination of correlation between cell strength and keratin gene mutations in human keratinocytes |
| Sekundarni povzetek: |
Keratin filaments are a major structural element in epithelial cells. They maintain shape and stiffness of cells by providing physical resistance to mechanical stress. Several different mutations in keratins leading to different types of disease have been described. Mutations in keratins 5 and 14 are known to cause the formation of epidermolysis bullosa simplex (EBS), a disease of skin fragility. In the master's thesis, we studied the point mutation K14 R125P, which causes a severe form of EBS. A typical phenotypic phenomenon with mutation is the formation of aggregates, which has not yet been explained in detail. Using optical tweezers, the stiffness of keratinocytes was determined by inserting a construct with additional copy of K14, with the R125P mutation, for different amounts of mutated keratin. We found that the difference in stiffness occurs only in cells with the lowest amount of mutated keratin, where we also observed the most aggregates. Aggregates are a dynamic organelle that can be recycled by a cell through a proteosomal system, as demonstrated by inhibition and western transfer. With mathematical modeling we also explained the formation of aggregates - with asymmetric binding in which both wild-type keratin and mutated keratin participate, and later the aggregates grow with the addition of mainly mutated keratin. |
| Sekundarne ključne besede: |
epidermis;keratinocytes;bullous epidermolysis simplex;keratin 14;aggregates; |
| Vrsta dela (COBISS): |
Magistrsko delo/naloga |
| Študijski program: |
1000377 |
| Konec prepovedi (OpenAIRE): |
1970-01-01 |
| Komentar na gradivo: |
Univ. v Ljubljani, Fak. za kemijo in kemijsko tehnologijo, smer Biokemija |
| Strani: |
53 str. |
| ID: |
13356516 |